Trovafloxacin-induced replication stress sensitizes HepG2 cells to tumor necrosis factor-alpha-induced cytotoxicity mediated by extracellular signal-regulated kinase and ataxia telangiectasia and Rad3-related

Abstract

Use of the fluoroquinolone antibiotic trovafloxacin (TVX) was restricted due to idiosyncratic, drug-induced liver injury (IDILI). Previous studies demonstrated that tumor necrosis factor-alpha (TNF) and TVX interact to cause death of hepatocytes in vitro that was associated with prolonged activation of c-Jun N-terminal kinase (JNK), activation of caspases 9 and 3, and DNA damage. The purpose of this study was to explore further the mechanism by which TVX interacts with TNF to cause cytotoxicity. Treatment with TVX caused cell cycle arrest, enhanced expression of p21 and impaired proliferation, but cell death only occurred after cotreatment with TVX and TNF. Cell death involved activation of extracellular signal-related kinase (ERK), which in turn activated caspase 3 and ataxia telangiectasia and Rad3-related (ATR), both of which contributed to cytotoxicity. Cotreatment of HepG2 cells with TVX and TNF caused double-strand breaks in DNA, and ERK contributed to this effect. Inhibition of caspase activity abolished the DNA strand breaks. The data suggest a complex interaction of TVX and TNF in which TVX causes replication stress, and the downstream effects are exacerbated by TNF, leading to hepatocellular death. These results raise the possibility that IDILI from TVX results from MAPK and ATR activation in hepatocytes initiated by interaction of cytokine signaling with drug-induced replication stress.

Tissue influx of neutrophils and monocytes is delayed during development of trovafloxacin‐induced tumor necrosis factor‐dependent liver injury in mice

“…We also demonstrate that TVX has clear effects before TNF injection, i.e., low level of DNA fragmentation in liver and reduced amounts of macrophages in the spleen. It is known that TVX by itself induces compound‐specific effects in hepatocytes (such as DNA damage caused by topoisomerase II inhibition) (Beggs et al, ; Beggs, Fullerton, Miyakawa, Ganey, & Roth, ; Liguori, Blomme, & Waring, ), although the cytotoxic (i.e., apoptosis) effects of TVX in vitro also depend on the presence of TNF (Beggs et al, ; Cosgrove et al, ). However, we cannot exclude that low levels of TNF, e.g., induced by subtle leakage of LPS from the gut, are involved in these effects.…”

Endoplasmic Reticulum Stress Induction and ERK1/2 Activation Contribute to Nefazodone-Induced Toxicity in Hepatic Cells

“…Recently published studies suggest that by interacting with various signaling pathways, the MAPK signaling cascades play pivotal roles in drug-induced liver toxicity (Beggs et al ., 2015; Chen et al ., 2014d, 2015; Hu et al ., 2016; Poulsen et al ., 2014; Wu et al ., 2015). The MAPK cascade has 3 well-defined arms: extracellular signal-regulated kinase 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 (Pearson et al ., 2001).…”

Idiosyncratic Drug-Induced Liver Injury: Is Drug-Cytokine Interaction the Linchpin?

“…TNFa potentiated cytotoxicity of chlorpromazine in primary mouse hepatocytes via activation of JNK (Gandhi et al, 2010). TVX synergized with TNFa to cause cell death in HepG2 cells and primary mouse hepatocytes (Beggs et al, 2015). The TVX/TNFa-induced cytotoxic interaction depended on the MAPKs JNK and ERK and also on ataxia telangiectasia Rad3-related, which is activated in response to replication stress and DNA damage (Beggs et al, 2014(Beggs et al, , 2015.…”

“…Moreover, numerous drugs that cause human IDILI inhibit cell proliferation in vitro. Examples include diclofenac (Rajabalian et al, 2009), sulindac (Chennamaneni et al, 2012), TVX (Beggs et al, 2015), halothane (Waxler et al, 1994), DOX (Supino et al, 1977), and chlorpromazine (Basta-Kaim et al, 2006). In addition, a series of NSAIDs associated with IDILI inhibited HepG2 cell proliferation in vitro, whereas an NSAID not associated with IDILI, aspirin, did not have this effect (Maiuri et al, 2014).…”